{"id":1296,"date":"2014-03-04T10:18:10","date_gmt":"2014-03-04T15:18:10","guid":{"rendered":"https:\/\/www.bowdish.ca\/lab\/?page_id=1296"},"modified":"2018-12-29T10:15:05","modified_gmt":"2018-12-29T15:15:05","slug":"streptococcus-pneumoniae","status":"publish","type":"page","link":"https:\/\/www.bowdish.ca\/lab\/protocols-old\/streptococcus-pneumoniae\/","title":{"rendered":"Streptococcus pneumoniae (old)"},"content":{"rendered":"<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2014\/03\/Culturing-Streptococcus-pneumoniae.pdf\">Growth and preparation of\u00a0<em>Streptococcus pneumoniae<\/em><\/a><\/p>\n<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2013\/02\/Streptococcus-pneumoniae-colonization1.pdf\">Intranasal colonization of\u00a0<em>Streptococcus pneumoniae<\/em><\/a><\/p>\n<ul>\n<li>This is a detailed protocol for preparing\u00a0<em>Streptococcus pneumoniae\u00a0<\/em>and intranasal colonization of mice.<\/li>\n<\/ul>\n<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/07\/CFSE-labeling-of-S-pneumoniae.pdf\">CFSE labelling of<em>\u00a0S pneumoniae<\/em><\/a><\/p>\n<ul>\n<li>This protocol uses CFSE to label\u00a0<em>S. pneumoniae<\/em>\u00a0for flow cytometry or microscopy experiments.<\/li>\n<\/ul>\n<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/07\/NF-KB-Luciferase-Assay-Protocolv2.pdf\">NF-KB Luciferase Assay using HEK293T cells<\/a><\/p>\n<ul>\n<li>This protocol can be used to test whether bacterial ligands induce activation of TLRs and NLRs.<\/li>\n<\/ul>\n<p><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/07\/Macrophage-Killing-Assay.pdf\">Macrophage Killing Assay<\/a>\u00a0version 1<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2014\/03\/Macrophage-Killing-Assay.pdf\">Macrophage Killing Assay<\/a>\u00a0version 2<\/p>\n<ul>\n<li>How do you measure how well macrophages kill bacteria (or whether your treatment alters killing)? This is a standard kill curve protocol and a workhorse of the Bowdish lab. Specifically written to measure\u00a0<em>S. pneumoniae<\/em>\u00a0killing, can be adapted for any bacteria and any type of macrophage (e.g. human, murine, alveolar, bone marrow,etc).<\/li>\n<\/ul>\n<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/08\/Pneumoncoccal-Cell-Wall-Purification-2DB-edits.pdf\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2016\/05\/Pneumococcal-Cell-Wall-Purification.pdf\">Pneumococcal Cell Wall Purification<\/a><\/p>\n<ul>\n<li>This protocol describes how to purify the pneumococcal cell wall. The purified preparation will not contain proteins but will contain the peptidoglycan layer and teichoic acids. It is updated from the <a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/08\/Pneumoncoccal-Cell-Wall-Purification-2DB-edits.pdf\">original<\/a>.<\/li>\n<\/ul>\n<p><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/08\/Pneumoncoccal-Cell-Wall-Purification-2DB-edits.pdf\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2016\/05\/Mannose-Detection-Protocol.pdf\">Phenol-sulfuric acid Mannose detection assay<\/a>.<\/p>\n<ul>\n<li>This assay is sensitive to mannose from 1 \u2013 100 ?g but will also detect\u00a0other carbohydrates. We use this assay to\u00a0quantify of complex carbohydrates in the bacterial cell wall<\/li>\n<\/ul>\n<div><a href=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2012\/08\/Pneumoncoccal-Cell-Wall-Purification-2DB-edits.pdf\"><img loading=\"lazy\" decoding=\"async\" title=\"Macrophage-bullet\" src=\"https:\/\/www.bowdish.ca\/lab\/wp-content\/uploads\/2011\/07\/Macrophage-bullet.gif\" alt=\"\" width=\"20\" height=\"20\" \/><\/a>\u00a0Nasopharyngeal colonization, and measurement of RNA and leukocyte recruitment.<\/div>\n<div>For a detailed protocol, see our paper here&#8230;<\/div>\n<div>\n<p>Puchta A, Verschoor CP, Thurn T, Bowdish DM.\u00a0<a href=\"http:\/\/www.jove.com\/video\/50490\/characterization-inflammatory-responses-during-intranasal\">Characterization of inflammatory responses during intranasal colonization with <\/a><em><a href=\"http:\/\/www.jove.com\/video\/50490\/characterization-inflammatory-responses-during-intranasal\">Streptococcus pneumoniae.<\/a>\u00a0<\/em>J Vis Exp. 2014 Jan 17;(83):e50490. doi: 10.3791\/50490.<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Growth and preparation of\u00a0Streptococcus pneumoniae Intranasal colonization of\u00a0Streptococcus pneumoniae This is a detailed protocol for preparing\u00a0Streptococcus pneumoniae\u00a0and intranasal colonization of mice. CFSE labelling of\u00a0S pneumoniae This protocol uses CFSE to label\u00a0S. pneumoniae\u00a0for flow cytometry or microscopy experiments. NF-KB Luciferase Assay &hellip; <a href=\"https:\/\/www.bowdish.ca\/lab\/protocols-old\/streptococcus-pneumoniae\/\">Continue reading <span class=\"meta-nav\">&rarr;<\/span><\/a><\/p>\n","protected":false},"author":2,"featured_media":0,"parent":436,"menu_order":0,"comment_status":"open","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-1296","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/pages\/1296","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/comments?post=1296"}],"version-history":[{"count":10,"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/pages\/1296\/revisions"}],"predecessor-version":[{"id":2715,"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/pages\/1296\/revisions\/2715"}],"up":[{"embeddable":true,"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/pages\/436"}],"wp:attachment":[{"href":"https:\/\/www.bowdish.ca\/lab\/wp-json\/wp\/v2\/media?parent=1296"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}